Week 3

Question 1

1.In gel electrophoresis, how much agarose does a 2% gel have (if the total gel makes up 40ml)?

Answer 1

0.8g
1g = 1ml So 2% of 40ml is 0.02*40 = 0.8g

Question 2

2.Which components are we looking to remove from our sample by performing PCR purification?

Answer 2

Primers
Taq polymerase
Contaminats

We only want to leave our purified PCR product for the next process.

Question 3

What charge does the silica matrix have in the spin column which allows DNA to bind to it? Tick all that apply.

Answer 3

Positive as DNA is negative.

Question 4

4.Which type of nucleotide is responsible for terminating chain synthesis in Sanger sequencing?

Answer 4

ddNTP- Dideoxynucleoside triphosphates lack the -OH group at the 3’ carbon, so no more bases can be added to the chain

Question 5

5.How many primers do we add to each cycle sequencing reaction?

Answer 5

1- If we added more we’d have multiple chains being synthesised at once which would make interpreting the sequence impossible with Sanger sequencing.